Assessment of Sanitation
Practices
Michael S. Ramsey
Teaching Laboratory Manager
UCD
mramsey@ucdavis.edu
• Talk today assumes risk assessment and
design and implementation of Winery
Sanitation and Quality Program
• The link to my talk is still up but brutal to
list
• Google “Michael Ramsey” and “HACCP”
Objectives
• Discuss “Quality”
• Methods to Assess Sanitation Practices
– Other industries
– Non-microbiological
– Microbiological
• Cell swabbings
• ATP Bioluminescence
What is Wine Quality?
• “Quality... you know what it is, yet you
don’t know what it is.”
– Robert Pirsig (1974); Zen and the Art of
Motorcycle Maintenance; pg 178
What is Wine Quality
• All sectors of manufacturing debate the definition
of “quality”
• To the ancient Greeks, “quality” meant “truth”
– They would not have understood our usage of the
word in our modern wine making dilemma
– Aristotle said “Quality (Truth) is not an act, it is a
habit”
• The Oxford English Dictionary (OED) defines
“quality”:
– “the degree of excellence of the thing”
What is Wine Quality?
• Why isn’t wine quality
as easy to define as,
say, golf ball quality?
“You can’t fake quality
any more than you
can fake a good
meal”
- William S. Burroughs
What is Wine Quality?
• Example: the ester, ethyl acetate
– Produced by some microbes that can be present
during wine fermentation
– This character is found at high concentration in wine
vinegar
– It is also a principle aroma compound of many nail
polish removers.
• People who associate this character with nail
polish remover tend to consider it quite
objectionable in wines
• People who associate it with vinegar do not find
it as unpalatable
What is Wine Quality?
• As Pirsig implied, we all have different
understandings of “quality” and that these
understandings are inherently subjective
What is Wine Quality?
• In other manufacturing sectors, “quality” is about
minimizing variability and meeting clear
specifications – or “zero defects”
• OED definition of defect: a shortcoming,
imperfection, or lack
• Origin: late Middle English (as a noun,
influenced by Old French defect 'deficiency'):
from Latin defectus, past participle of deficere
'desert or fail', from de- (expressing reversal) +
facere 'do'
What is Wine Quality?
• If “quality” means “zero defects” then a defective
wine is one that is purchased by your consumer
in any condition other than how you intended for
it to be in the bottle.
• Wine “Quality” is ultimately how you – or your
customer - define it.
“Quality means doing it right when no one is
looking.”
– Henry Ford
• How does “quality”, then, impact one’s
assessment of my sanitation practices?
Questions to Ask Yourself
• What should be the main emphasis of my
definition of quality?
• What are the important areas of risk and
sanitation in my winery to look into and
why?
• How much risk am I willing to accept?
• Which method(s) of assessment are most
suitable for my vision of Quality and
Quality Assurance?
• Traditionally, a combination of three methods
are used to assess efficacy of sanitation
methods
– Education and Training
– Inspections (visual assessments)
– Microbiological testing
• “We are stuck with technology when what we
really want is just stuff that works.”
– Douglas Adams, The Salmon of Doubt (2002)
Education and Training
• Requires continuous work due to
employee turnover
– Which is rarely done
Visual Assessments
• Limited
• Often, a lack of specificity (terms like
satisfactory, acceptable, etc.) leaves the
interpretation to the inspector, who may
place too much emphasis on relatively
unimportant matters and thus increase
costs without reducing hazards.
• Not always reliable to detect sanitary
conditions
Microbiological Methods of
Assessment in Food
• Molding surfaces with agar
• Staining surfaces and image analysis with
epifluorescence microscope
• Bacterial impedance (i.e., BacTrac 4300)
– A sample is incubated and tested for changes
in impedance in a specific medium
Microbial Methods in Wineries
• Microbial Swabbing
• ATP Bioluminescence
Microbiological Swabbing
• Apply sterile swab
over 4 in. x 4 in.
surface area
• Swab is transferred to
sterile liquid &
vortexed
• Liquid is put on
selective plates which
allow microbes to
grow
Microbiological Swabs
Microbiological Swabs
• Positive attributes
– Possible to quantify number and type of microbes
present
– Can be used to trend sanitary condition of
facility/equipment.
• Technology presumes there's just one right way
to do things and there never is.
– Robert M. Pirsig, Zen and the Art of Motorcycle
Maintenance (1974), p. 166
Microbiological Swabs
• Microbiological testing also has some
limitations
– Results not available for days
– TVC (Total Viable Counts) is not really “total”.
It is only a measure of the fraction of the
microbes able to produce colonies in the
medium used under the conditions of
incubation.
• Difficulties related to sampling
– Training, swab material, and technique
ATP Bioluminescence
• Widespread use in dairy, food, and beer
industries since the early 1990s
• ATP is short for adenosine triphosphate.
– This compound is the basic currency for
energy in all living cells.
ATP Bioluminescence
• The presence of this
molecule may be
assayed simply using
an enzyme and
coenzyme complex
(luciferase/luciferin)
found in the tail of the
firefly, Photinus
pyralis.
ATP Bioluminescence
• The reaction is essentially the stoichiometric
conversion of ATP to photons of light in the
presence of the enzyme luciferase
ATP Bioluminescence
• luciferin + ATP + O2 + Mg2+
oxyluciferin + AMP + PPi + CO2 + light
• The breakthrough idea was to use ATP
from all biological residues on a surface
as an indicator of cleanliness, rather than
try to ascertain the number of
microorganisms present.
ATP Bioluminescence
• The total ATP present on a swabbed surface
can be extracted, assayed, and potentially
uploaded to a computer database extremely
rapidly (within five minutes).
ATP Bioluminescence
• The light given off in this
reaction is assayed as
Relative Light Units
(RLUs) – a measure of
the light produced by the
ATPase assay relative to
a control sample.
• RLU readings above the
number you have chosen
(based on the sum of
your interferences) would
be “FAIL”events in your
cleaning/sanitation
program.
ATP Bioluminescence
• Since the results are “relative”, we can not
compare results from one system to
another.
• However, these assays are sensitive,
direct, objective tests of cleaning efficiency
and risk.
ATP Bioluminescence
• Although the ATP bioluminescence assay is very
rapid, it is not very specific.
– The results tell us nothing about the nature of the
contaminant, whether it is juice, wine, random organic
matter, microorganisms or combinations.
• There is no direct correlation between ATP
results and microbial counts:
• ATP amounts can vary widely between
microbes.
ATP Bioluminescence
• If your QA strategy is to provide quick and
sensitive answers to your winery cleaning
efficiency, or your bottling line sanitation
requires “zero” contaminants regardless of
the nature of that contamination, ATP
Bioluminescence assays might just be for
you.
Issues With Both Methods
• Cell culture and ATP methods tend to under count
remaining cells
• Training and technique
• There is a limit of detection with swabbing
– Depending on area swabbed and swab material
• All ATP-Bioluminescence systems also have a limit of
detection
– No system can presently detect 100% of all ATP present.
• The lower the limit of detection, the more sensitive a
system will be. Generally, LODs decrease as the price of
the unit increases.
• You will need to decide how much sensitivity fits into
your vision of quality and your budget.
Conclusions
• A combination of three methods are used to
assess efficacy of sanitation methods
– Education and Training
– Inspections (visual assessments)
– Microbiological testing
• For further discussion of ATP methods in the
winery, refer to Ramsey, M. S. and Mills, D.
A. 2013. Winery Biofilms: A source for
contamination throughout the winery. Practical
Winery and Vineyard Journal. Winter Issue.
Thank you
• “Making an art out of your technological
life is the way to solve the problem of
technology.”
– Robert Pirsig,1974